BACKGROUND:Up to date,inbred embryonic stem cells(ESCs) line mainly derived from 129 mouse strain and C57BL/6 strain,occasionally from BALB/c mouse strain,however,few reports concerning ESCs lines derived from Ch inese Kunmingmouse strain.OBJECTIVE:To isolate and culture mouse ESCs of kunming species,additionally,to identify its biological properties.DESIGN,TIME AND SETTING:The experiment with cells as observed subjects was conducted in the institute of Urology,First Affiliated Hospital of Nanchang University from May 2006 to June 2007.MATERIALS:Blastocysts of Kunmin mice with 3.5 days of embryonic age.METHODS:Inner cell mass from 3.5 days embryonic age were isolated from Kunming species mice,cultured on feeder layers of primary mice embryonic fibroblasts,and then the cells were isolated and subsequently cultured.MAIN OUTCOME MEASURES:Colony growth was observed and determined by alkaline phosphatase (AKP) staining;The expression of stage-specific embryonic antigen (SSEA)-1,and cell specific genes of c-Myc,Nanog,Oct3/4 and Sox2 were measured by immunofluorescence and RT-PCR;finally,the ability of differentiation in vitro and in vivo was identified.RESULTS:The ESCs-like colonies presented typical morphological characteristics of ESCs,which was positive to AKP and SSEA-1,and could express several cell specific genes,such as c-Myc,Nanog,Oct3/4 and Sox2,and differentiate into various cell types in vitro and in vivo.CONCLUSION:An ESCs line is successfully dedved from Kunming mice,which has typical biological characteristics of ESCs.

Objective To investigate the expression changes of microRNAs and VEGF-NOTCH in renal ischemic injury in mice, and to explore the potential mechanism associated with renal angiogenesis.Method Male Balb/c mice were subjected to a standard renal ischemia to induce acute kidney injury (AKI) after 45 min of bilateral renal artery clamping. Following 4 h, 24 h of reperfusion or sham operation, kindey tissues were collected and subjected to detect the expression changes of microRNAs which relatived with angiogenesis and VEGF, Flk-1, Notch1 mRNA by Quantitative Real-time RT-PCR. Flk-1 protein was detected by Western blotting analysis at 24 h and 72 h following Ischemia/Reperfusion(I/R) injury. The expression of CD31 was examined in tissue sections by immunohistochemistry staining, and the microvessels in ischemic region of each group were counted. Results miRNA-210 and miRNA-92a expression increased significantly, with prominent changes at 4 h and 24 h after reperfusion( P ＜ 0.05 ). VEGF and Flk-1 mRNA expression and Flk-1 protein were increased in renal I/R compared with control group respectively (P＜0.05 ).Immunohistochemistry staining results of CD31 showed a significant increase of microvessels in renal ischemic region. Conclusion This study first reported the changes in miRNAs expression in response to kidney I/R in mouse. our results implied that miRNAs may be involved in targeting VEGF-Notch pathway signaling to regulate angiogenesis after renal I/R injury. It provided novel insights into the angiogenesis mechanism of renal ischemic injury.

Immuno-fluorescence technique can qualitatively determine certain nuclear translocation, of which NF-κB/ p65 implicates the activation of NF-κB signal pathways. Immuno-fluorescence analysis software with independent property rights is able to quantitatively analyze dynamic location of NF-κB/p65 by computing relative fluorescence units in nuclei and cytoplasm. We verified the quantitative analysis by Western Blot. When we applied the software to analysis of nuclear translocation in lipopolysaccharide (LPS) induced (0. 5 h, 1 h, 2 h, 4 h) primary human umbilical vein endothelial cells (HUVECs) , we found that nuclear translocation peak showed up at 2h as with calculated Western blot verification results, indicating that the inventive immuno-fluorescence analysis software can be applied to the quantitative analysis of immuno-fluorescence.
Active Transport, Cell Nucleus ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Fluorescent Antibody Technique ; Human Umbilical Vein Endothelial Cells ; Humans ; NF-kappa B p50 Subunit ; metabolism ; Software

ObjectiveTostudythemolecularmechanismoflanthanumonblocking lipopolysaccharide(LPS)-mediated activation of nuclear factor-kappaB (NF-κB)signaling in macrophages.MethodsThe RAW264.7 macrophages were cultured routinely and divided into 4 groups randomly:LaCl3 +LPS group,LPS group,LaCl3 group and control group.The nuclear translocation of p65 protein was detected by immunocytochemistry.Total,cytoplasmic and nuclear proteins were extracted respectively,and then the binding activity of p65 with the target gene was measured by ELISA.Western blot assays were also performed to detect the expression levels of the proteins,including nuclear p65,IκBα and IKK kinase,the phosphorylation status of IκBα and IKK kinase.ResultsLanthanum can block LPS-induced activation of p65 protein through various ways,such as inhibiting its nuclear translocation,reducing its expression in the nuclei and decreasing its binding activity with the target genes.Lanthanum inhibited the degradation of LPS-induced IκBα,but the phosphorylation of LPS-induced IKKβ can not be blocked by lanthanum,nor did the phosphorylation ability of p-IKKβ on IκBα.Conclusion Lanthanum inhibited the degradation of IκBα,nuclei translocation of p65 protein and its binding activity with the target genes,thereby inhibited LPS-induced NFκB activation,which might be one of the inhibition mechanisms of lanthanum on nuclear activation.

In previous studies, four endophytic fungi were isolated from different swollen roots of Rehmannia glutinosa. It's thought that Ceratobasidium sp. , one of the discovered endophytic fingi, was a major promoter for the growth of the roots. In this study, symbiotic experiments were performed to measure the effects of different endophytic fingi cultivated with R. glutinosa. The results showed that the R. glutinosa had significant increases in the size of roots and amount of chlorophyll cultivated with Ceratobasidium sp. And it was tested that indoleacetic acid secreted by Ceratobasidium sp. maybe the effective factor for the promotion of the growth.
Fungi ; metabolism ; physiology ; Indoleacetic Acids ; metabolism ; Plant Roots ; growth & development ; microbiology ; Rehmannia ; growth & development ; microbiology ; Symbiosis ; physiology

Purpose To explore the application of vascular echo tracking technology in early stage vascular lesions of type 2 diabetes mellitus combined with proteinuria with normal intima and medial membrane of femoral artery,in order to provide evidence for early clinical intervention and to reduce complications of diabetes patients.Materials and Methods Fifty-two patients with type 2 diabetes mellitus diagnosed in Longgang Central Hospital from March 2015 to July 2016 were selected,20in proteinuria group and 32 in group without proteinuria.30 patients at the same stage with normal physical examination results were selected as control group.Vascular echo-tracking technology was used to check femoral arterial elasticity on both sides of the examinee,including stiffness parameter (β),pressure-strain elasticity modulus (Ep),arterial compliance (AC),augmentation index (AI) and one-point pulse wave velocity (PWVβ).Results ① Ep,β and PWVβ of femoral artery in diabetes patients were higher than those in control group,and AC was lower than that in control group.The differences was of statistical significance (P＜0.05).There was no statistical difference in AI between the two groups (P＞0.05).② Ep,β and PWVβ of femoral artery for patients in proteinuria group were higher than those in group without proteinuria and AC was also lower.The difference was of statistical significance (P＜0.05).There was no statistical difference in AI between the two groups (P＞0.05).③ After judgment and analysis,AC and diastolic blood pressure were of statistical significance for judgment,and sensitivity was 92.3％ and specificity 100.0％ when adopting its function to diagnose diabetes femoral arterial elasticity.Conclusion Vascular echo-tracking technology can accurately assess femoral elasticity for diabetes patients and is of great significance in early preventive treatment of diseases.The appearance ofproteinuria indicates that vascular elasticity function decreases dramatically.

Objective:To investigate the effect of extracellular vesicles derived from lung tissue on intrapulmonary inflammation and the formation of neutrophil extracellular traps (NETs) in sepsis rats.Methods:Sepsis rat model was established by cecal ligation and puncture (CLP). Collagenase D and DNase I were used to dissociate the lung tissue, the impurities were removed by centrifugation, and finally, the extracellular vesicles (Ti-EVs) derived from lung tissue were separated and extracted by differential ultracentrifugation. Eighteen male SD rats were randomly divided into the sham group, sepsis group and Ti-EVs group: in the Ti-sEV group, a sepsis model was established by CLP, and Ti-EVs suspension was instilled through the airway; rats in the CLP group received CLP, and an equal volume of PBS was instilled through the airway; and rats in the sham group was treated with sham operation. The pathological changes of lung tissue were detected by hematoxylin-eosin (HE) staining after 24 h. The content of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the bronchoalveolar lavage fluid (BALF) was measured by enzyme-linked immunosorbent assay (ELISA). Immunofluorescence was used to detect the NETs content in lung tissue.Results:The isolated extracellular vesicles derived from rat lung tissue were observed by transmission electron microscopy as double-layer circular cystic vesicles with particle diameter mainly distributed at 150 nm. Western blot showed positive expression of EVs markers CD9, CD63, and Tsg101. HE staining of lung tissue showed alveolar integrity damage and a large number of inflammatory cells infiltrated in the lung of sepsis rats. Compared with the CLP group, the degree of lung tissue damage was more serious in the Ti-EVs group and the levels of IL-1β, TNF-α and IL-6 in the BALF of rats were significantly increased ( P<0.01). The formation of NETs in the lungs of the rats in the sepsis group and the Ti-EVs group was observed under the laser confocal microscope. Compared with the sepsis group, the fluorescence intensity of NETs in the lung tissues of the Ti-EVs group increased significantly. Conclusions:After enzymatic digestion, differential ultracentrifugation and other treatments, the extracellular vesicles derived from rat lung tissue with high purity can be successfully isolated and extracted. In the process of septic lung injury, extracellular vesicles in lung tissue can aggravate the inflammatory response in the lung and promote the formation of NETs.

Major infectious disease epidemic continues to pose a threat to human health and society, and the effective establishment and implementation of an early warning system plays a key role in addressing public health security risks. At present, the research on early warning of infectious diseases in the academic community mainly focuses on early warning information system, early warning mechanism, laws and regulations of early warning of infectious diseases, and some studies lack specific suggestions on operation methods. By collating and summarizing the literature from 2002 to 2022, regarding the early warning system and mechanism of major infectious diseases, this paper focused on analyzing the public health ethical dilemmas existing in the early warning process and discussing how to strengthen the construction of the early warning system of infectious diseases, so as to lay the foundation for creating more scientific early warning schemes of infectious diseases.

Traditional Chinese medicine （TCM） has special advantages and independent value in fighting major epidemics such as emerging infectious diseases. To exert the efficacy of TCM in the field of public health emergency management， it is necessary to clarify the necessity and analyze the obstacles， hidden dangers， and ethical risks of integrating TCM into the public health emergency management system. By optimizing the legal system of integrating TCM into the public health emergency management system， improving the construction of system and mechanism of health emergency management for preventing and controlling major epidemics with TCM， enhancing the strength of clinical scientific research technology of TCM in preventing and controlling epidemics， strengthening the discipline education of TCM and the construction of the talent team， based on the background of combining Chinese and Western medicine， the integration of TCM into the national public health emergency management system can be promoted.